1. Conditional Immortalization Establishes a Repertoire of Mouse Melanocyte Progenitors with Distinct Melanogenic Differentiation Potential 
Ke Yang, Jin Chen, Wei Jiang, Enyi Huang, Jing Cui, Stephanie H. Kim, Ning Hu, Hong Liu, Wenwen Zhang, Ruidong Li, Xiang Chen, Yuhan Kong, Jiye Zhang, Jinhua Wang, Linyuan Wang, Jikun Shen, Hue H. Luu, Rex C. Haydon, Xiaohua Lian, Tian Yang, Tong-Chuan He 
Journal of Investigative Dermatology 
Volume 132, Issue 10, Pages (October 2012)
DOI: /jid
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2. Figure 1
Characterization of SV40T-immortalized melanocytes (iMCs). (a) iMC23 (aA) and iMC65 (aB) were seeded at low density and photographed at day 5. (bA) iMCs were infected with AdGFP or AdFLP. Green fluorescent protein signal was detected at 24hours. (bB) iMCs were infected with AdFLP (+) or AdGFP (-) for 48hours. Total cellular proteins were analyzed by anti-SV40T western blotting (β-actin blotting for loading). (bC, bD) iMC23 cells were infected with AdFLP or AdGFP, stained with Crystal violet, and the absorbance was quantitatively measured at 560nm. (cA) iMCs were collected by pelleting. Pigmentation of cell pellets was photographed. (cB) iMCs were collected for endogenous tyrosinase enzymatic assays. (dA) iMCs were immunostained with c-kit and HMB45 antibodies. (dB) iMCs were subjected to Fontana–Masson staining of melanin (arrows). OD, optical density. Bar=50μm.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
Melanogenic potential of immortalized melanocytes (iMCs). (aA) iMC23 cells were treated with dexamethasone (Dex) for 5 days and collected for tyrosinase enzymatic assay. (aB) iMC23 was stimulated with 12nM Dex. Gene expression was determined by reverse transcriptase–PCR (RT-PCR; glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as control). (b) iMC23 was treated with 12nM Dex for 5 days. Melanin production was visualized (bA) in cell pellet, (bB) in culture under the microscope, or by (bC) Fontana–Masson staining. Melanin-positive cells are indicated with arrows. Bar=100μm. (cA) Luciferase-tagged iMC23 and iMC65 infected with AdFLP or AdGFP were injected subcutaneously into flanks of athymic mice (arrows) and subjected to Xenogen imaging. (cB) Visual pigmentation at injection sites (arrows). (d) Injected sites were retrieved at day 7 and subjected to (dA) HMB45 immunostaining (HRP-AEC) and (dB) Fontana–Masson staining. Melanin-positive cells are indicated with arrows. Bar=25μm.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions